Extraction and purification of brain glycoprotein NSA3 by binding with hyaluronic acid

Brain glycoprotein NSA3 was found to bind to immobilised hyaluronic acid. The binding was reversible and gave pure antigen (99%) in high yields (80%). The binding was suppressed by incubating the affinosorbent with hyaluronidase. It was not suppressed by trypsin. The presence of glycosaminoglycans other than hyaluronic acid in the sample did not inhibit the binding. This property is relevant to those already known for the brain glycoprotein NSA3, and to its localisation at the nodes of Ranvier. We propose to coin the name hyaluronectin for the brain glycoprotein NSA3.     

NSA2在激光诱导的小鼠脉络膜新生血管模型中表达的时相性和定位研究

目的探讨NSA2在激光诱导的小鼠脉络膜新生血管（choroidal neovascularization，CNV）模型眼组织中的表达及其意义。方法532nm激光诱导C57BI/6J小鼠CNV模型。用CD31抗体标记血管内皮细胞的方法对CNV模型进行鉴定，采用RT-PCR和Western blotting方法检测正常对照组和光凝后1d、3d、5d、7d和14d组小鼠CNV中NSA2mRNA和蛋白表达的时间变化规律。取光凝后7d组小鼠眼做眼球冰冻切片，用免疫荧光染色方法对NSA2蛋白在CNV中的表达进行定位研究。结果NSA2在正常小鼠的视网膜脉络膜组织中弱表达。视网膜光凝后，NSA2mRNA和蛋白在CNV模型眼组织中的表达有明显的时间变化规律，光凝后l～3d逐渐增强，3d达高峰，之后逐渐减弱，14d时仍稍高于正常水平。同时发现NSA2在CNV区域表达较强。结论NSA2在CNV形成早期表达上调，具有明显的时间变化规律，且在CNV区域有较强的表达，因此我们推断NSA2可能在CNV形成这一病理过程中起着重要作用。     

Effects of monochromatic X-radiation on the membrane of nodes of Ranvier under voltage and current clamp conditions.

Monochromatic Ag-Ka-radiation decreased irreversibly the peak sodium current in nodes of Ranvier. This decrease occurs only with a delay of about 1000 sec after a threshold dose of about 8 kR has been reached. Potassium current and resting potential are practically not affected.     

Effects of monochromatic X-radiation on the membrane of nodes of ranvier under voltage and current clamp conditions

Summary Monochromatic Ag-K_a-radiation decreased irreversibly the peak sodium current in nodes of Ranvier. This decrease occurs only with a delay of about 1000 sec after a threshold dose of about 8 kR has been reached. Potassium current and resting potential are practically not affected.Dedicated to Prof. Dr Dres h.c. Robert Stämpfli on the occasion of his 65th birthday.Supported by Deutsche Forschungsgemeinschaft, SFB 38, Membranforschung Bonn-Bad Godesberg.     

Peripheral neuropathy associated with inhalation of methyl-n-butyl ketone

Résumé Dix-sept rats sont exposés pendant 6 semaines 5 jours par semaine, 8 h par jour, à une atmosphère contenant soit du méthyl-n-butyl kétone seul soit un mélange de MnBK et de méthyl-éthyl kétone. Tous les rats présentèrent une faiblesse musculaire généralisée après l'inhalation qui dura de quelques h à 24 h avec récupération motrice totale. En dépit de cette apparence normale, l'examen histologique révéla une hypertrophie, un ballonement en grains de chapelet, et une dégénérescence des axones, associée à une démyélination secondaire, habituellement située dans la région des nodes de Ranvier. La toxicité du MnBK est prouvée, celle du MEK est encore à l'étude.     

Elemental distribution of Na, P, Cl and K in different structures of myelinated nerve of Rana esculenta.

Measurements of the distribution of Na, P, Cl and K were performed in different structures of the myelinated nerve. Whereas the axon shows a typical intracellular distribution pattern for Na, Cl and K, the interstitial space and the myelin sheath show a typical extracellular pattern. These measurements have demonstrated that Na is present in the myelin sheath close to the node of Ranvier.     

Elemental distribution of Na,P, Cl and K in different structures of myelinated nerve ofRana esculenta

Summary Measurements of the distribution of Na, P, Cl and K were performed in different structures of the myelinated nerve. Whereas the axon shows a typical intracellular distribution pattern for Na, Cl and K, the interstitial space and the myelin sheath show a typical extracellular pattern. These measurements have demonstrated that Na is present in the myelin sheath close to the node of Ranvier.     

Selective blockade of components of potassium activation in Myxicola axons

The K+ conductance in Myxicola giant axons activates in two phases which are pharmacologically separable. The fast phase of K+ activation is specifically inhibited by 4-aminopyridine and by the substitution of D2O for H2O. We suggest Myxicola giant axons, like the amphibian node of Ranvier, may possess more than one variety of K+ channel.     

The effect of vinblastin and vincristin on single nerve fibres

Summary The perfusion of the node of Ranvier with vinblastin or vincristin reduces the amplitude of the action potential within a few seconds. Vincristins is 10fold more active than vinblastin. Upon withdrawal, the effect is promptly reversible and it is antagonized by acetylcholine.     

Intracellular localization of alpha-fetoprotein and serum albumin in the central nervous system of the rat during fetal and postnatal development]

The morphological localisation of alphafetoprotein (AFP), serumalbumin (SA), transferrin and immunoglobulins (IgG) has been studied in the developing central nervous system of the Rat by immunocytochemical methods. Evidence is presented of a highly selective staining for AFP and SA, both proteins exhibiting the same topographical distribution. Practically all the areas of the brain and the spinal cord are stained at a given moment of the developmental process. The labeling is cytoplasmic and in the neuronal elements extends to their axonic and dendritic prolongations. The localization of AFP and SA in the nervous system may be related to the well known binding properties of these proteins for varied substances (estrogen and/or fatty acids). The morphological data presented here suggest that both proteins may be actively involved in the uptake of such substances by the cellular structures of the nervous tissue.     

Starvation-induced changes in the autoradiographic localisation of valine uptake by rat small intestine

We report here the effects of a 72-h fast on the localisation of Na-dependent [3H]-valine uptake by rat small intestine. Starvation results in the earlier appearance of valine transport during cell migration and an enhanced accumulation of the amino acid at the villus tip.     

异丙酚对大鼠内毒素脑损伤STAT3表达的影响

目的 研究异丙酚对大鼠内毒素脑损伤中信号转导子与转录激活子-3( STAT3)表达的影响,探讨异丙酚在脑损伤中的保护作用及其机制.方法 健康清洁级SD大鼠72只,雌雄不限,体重220～ 250 g,随机分为3组(n=24):L组(内毒素组)和LP组(内毒素+异丙酚组)经颈内动脉注射内毒素200 μg建立大鼠内毒素脑损伤模型,C组(对照组)经颈内动脉注射等量生理盐水,LP组颈内动脉注射内毒素后即予异丙酚100 mg/kg剂量腹腔注射.3组分别于6、12、24和48h随机处死6只大鼠,取额叶皮质,检测脑组织含水量,免疫组织化学检测P-STAT3、NF-κB和iNOS表达水平的变化,Western blot法检测大鼠内毒素脑损伤后P-STAT3蛋白表达水平的变化.结果 与C组相比,L组、LP组各时间点脑组织含水量、P-STAT3、NF-κB和iNOS表达增加(P ＜0.05,P＜0.01);与L组比较,LP组各时间点脑含水量、P-STAT3、NF-κB和iNOS表达明显减少(P＜0.05,P＜0.01).结论 异丙酚可减轻大鼠内毒素性脑损伤,机制可能与抑制脑组织磷酸化STAT3、NF-κB和iNOS表达水平上调,进而减轻炎性反应有关.     

Two distinct phosphorylation events govern the function of muscle FHOD3

Posttranslational modifications such as phosphorylation are universally acknowledged regulators of protein function. Recently we characterised a striated muscle-specific isoform of the formin FHOD3 that displays distinct subcellular targeting and protein half-life compared to its non-muscle counterpart and which is dependent on phosphorylation by CK2 (formerly casein kinase 2). We now show that the two isoforms of FHOD3 are already expressed in the vertebrate embryonic heart. Analysis of CK2 alpha knockout mice showed that phosphorylation by CK2 is also required for proper targeting of muscle FHOD3 to the myofibrils in embryonic cardiomyocytes in situ. The localisation of muscle FHOD3 in the sarcomere varies depending on the maturation state, being either broader or restricted to the Z-disc proper in the adult heart. Following myofibril disassembly, such as that in dedifferentiating adult rat cardiomyocytes in culture, the expression of non-muscle FHOD3 is up-regulated, which is reversed once the myofibrils are reassembled. The shift in expression levels of different isoforms is accompanied by an increased co-localisation with p62, which is involved in autophagy, and affects the half-life of FHOD3. Phosphorylation of three amino acids in the C-terminus of FHOD3 by ROCK1 is sufficient for activation, which results in increased actin filament synthesis in cardiomyocytes and also a broader localisation pattern of FHOD3 in the myofibrils. ROCK1 can directly phosphorylate FHOD3, and FHOD3 seems to be the downstream mediator of the exaggerated actin filament formation phenotype that is induced in cardiomyocytes upon the overexpression of constitutively active ROCK1. We conclude that the expression of the muscle FHOD3 isoform is characteristic of the healthy mature heart and that two distinct phosphorylation events are crucial to regulate the activity of this isoform in thin filament assembly and maintenance.     

Selective blockade of components of potassium activation inMyxicola axons

Summary The K⁺ conductance inMyxicola giant axons activates in two phases which are pharmacologically separable. The fast phase of K⁺ activation is specifically inhibited by 4-aminopyridine and by the substitution of D₂O for H₂O. We suggestMyxicola giant axons, like the amphibian node of Ranvier, may possess more than one variety of K⁺ channel.     

求解椭圆方程的径向基无网格配置法

针对一、二维椭圆方程构造了径向基无网格配置法；给出了解的存在唯一性；同时得到了基函数中自由参数c与求解精度的关系，以及节点均布时自由参数最佳取值的计算公式。将节点均布下得到的自由参数取值公式应用于节点任意排列的情况，其求解精度仍能得到保证，表明这种无网格方法对节点的位置不敏感。     

Nucleolar localization of succinic dehydrogenase in human malignant cells with MTT

Résumé Les auteurs complètent leurs recherches sur la localisation cytochimique de l'activité de la déshydrogénase succinique dans les nucléoles en utilisant un troisième sel de tetrozolium, MTT d'une importance particulière.     

一种基于自体集层次聚类的否定选择算法

否定选择算法是用于产生人工免疫检测器的重要算法,然而传统的否定选择过程需要将随机生成的候选检测器与全部自体数据进行匹配以排除识别了自体的无效检测器,该匹配过程导致检测器的生成效率过低,极大地限制了免疫算法的应用.为此,文中提出了一种基于自体集层次聚类的否定选择算法CB-RNSA.算法首先对自体数据进行层次聚类预处理,然后用聚类中心取代自体数据点与候选检测器进行匹配,以减少距离计算代价.在生成检测器的过程中,候选检测器被限定在非自体空间的低覆盖率区域内,以降低检测器冗余.对检测器的非自体空间覆盖率进行了概率分析,给出了中止生成检测器的条件,该条件较传统的预设检测器数量的中止条件更为合理.理论分析表明CB-RNSA的时间复杂度与自体集规模无关,从而解决了经典的否定选择算法的时间复杂度随自体数量呈指数增长这一难题,极大地提高了大自体样本空间下的检测器生成效率.对比实验结果表明:在相同的实验数据集与期望覆盖率下,CB-RNSA的检测率比经典的RNSA与V-detector算法分别提高了12.3%与7.4%,误警率分别降低了8.5%与4.9%,产生检测器的时间代价分别降低了67.6%和75.7%.     

Neuronal ceroid lipofuscinosis protein CLN3 interacts with motor proteins and modifies location of late endosomal compartments

CLN3 is an endosomal/lysosomal transmembrane protein mutated in classical juvenile onset neuronal ceroid lipofuscinosis, a fatal inherited neurodegenerative lysosomal storage disorder. The function of CLN3 in endosomal/lysosomal events has remained elusive due to poor understanding of its interactions in these compartments. It has previously been shown that the localisation of late endosomal/lysosomal compartments is disturbed in cells expressing the most common disease-associated CLN3 mutant, CLN3?ex7-8 (c.462-677del). We report here that a protracted disease causing mutant, CLN3E295K, affects the properties of late endocytic compartments, since over-expression of the CLN3E295K mutant protein in HeLa cells induced relocalisation of Rab7 and a perinuclear clustering of late endosomes/lysosomes. In addition to the previously reported disturbances in the endocytic pathway, we now show that the anterograde transport of late endosomal/lysosomal compartments is affected in CLN3 deficiency. CLN3 interacted with motor components driving both plus and minus end microtubular trafficking: tubulin, dynactin, dynein and kinesin-2. Most importantly, CLN3 was found to interact directly with active, guanosine-5'-triphosphate (GTP)-bound Rab7 and with the Rab7-interacting lysosomal protein (RILP) that anchors the dynein motor. The data presented in this study provide novel insights into the role of CLN3 in late endosomal/lysosomal membrane transport.     

Glycogen synthase kinase 3: A key regulator of cellular fate

The serine/threonine kinase glycogen synthase kinase-3 (GSK-3) was initially identified as a key regulator of insulin-dependent glycogen synthesis. GSK-3 was subsequently shown to function in a wide range of cellular processes including differentiation, growth, motility and apoptosis. Aberrant regulation of GSK-3 has been implicated in a range of human pathologies including Alzheimer’s disease, non-insulin-dependent diabetes mellitus (NIDDM) and cancer. As a consequence, the regulation of GSK-3 and the therapeutic potential of GSK-3 inhibitors have become key areas of investigation. This review will focus on the mechanisms of GSK-3 regulation, with emphasis on modulation by upstream signals, control of substrate specificity and GSK-3 localisation. The details of these mechanisms will be discussed in the context of specific signalling pathways. Received 30 January 2007; received after revision 5 March 2007; accepted 16 April 2007     

Detection of a minor stimulating product involved in secondary allogenic proliferation of human lymphocytes in vitro]

In a family with a shared parental haplotype studied in MLR I and II we report that: 1) A secondary proliferation can be induced without a primary positive MLR; 2) In these conditions a minor determinant activating secondary proliferation is detected; 3) No significant association of this product with the available makers (HLA-A, B, C, D, Ly-Li) of the HLA region has been found so far; its localisation within or outside the MHC is under investigation.